Journal: Molecular Oncology

Article Title: Mechanistic insights into p53‐regulated cytotoxicity of combined entinostat and irinotecan against colorectal cancer cells

doi: 10.1002/1878-0261.13060

Figure Lengend Snippet: Impaired homologous repair after irinotecan plus entinostat combination in HCT116 wt cells. HCT116 wt and HCT116 Δp53 cells were exposed to 10 µ m irinotecan (Iri) ± 2 µ m entinostat (MS, short for MS‐275). Expression levels of indicated proteins were analyzed by immunoblot after 24 h (A) and 48 h (B). α‐tubulin was used to control protein loading. The mRNA expressions of indicated genes (C) were quantified by qRT‐PCR after 24 h. HCT116 wt (D) and HCT116 Δp53 (E) cells were exposed to 10 µ m Iri ± 2 µ m MS for 24 h or irradiated with 2 Gy for 2 h before fixation. The expression and localization of 53BP1 (green; upper panels) or RAD51 (green; lower panels) together with γH2AX (red) was analyzed by fluorescence microscopy. DAPI was used as a nuclear stain. Scale bars are equal to 10 µm. All graphs show the mean value of independent experiments ± SEM; immunoblots and microscopy pictures are representative of three independent experiments.

Article Snippet: Following, samples were incubated in primary antibody solutions against H2AX phosphorylated at serine‐139 (γH2AX; 1 : 1000 in BS; Merck Millipore), 53BP1 (1 : 500 in BS; Novus Biologicals), or RAD51 (1 : 250 in BS; Santa Cruz), washed in PBS and incubated in AlexaFluor 555‐conjugated anti‐rabbit (1 : 500 in BS; Abcam) or AlexaFluor 647‐conjugated anti‐mouse (1 : 500 in BS; Thermo Fisher) secondary antibody solutions.

Techniques: Expressing, Western Blot, Control, Quantitative RT-PCR, Irradiation, Fluorescence, Microscopy, Staining

Journal: Molecular Cell

Article Title: Ribonucleotide Reductase Requires Subunit Switching in Hypoxia to Maintain DNA Replication

doi: 10.1016/j.molcel.2017.03.005

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti-53BP1 , Novus Biologicals , Cat# NB100-904.

Techniques: Transduction, Recombinant, Protease Inhibitor, SYBR Green Assay, Mutagenesis, Purification, Gel Extraction, Imaging, Sequencing, Negative Control, Real-time Polymerase Chain Reaction, Software, Expressing

Journal: The Journal of Neuroscience

Article Title: Memory-Related Synaptic Plasticity Is Sexually Dimorphic in Rodent Hippocampus

doi: 10.1523/JNEUROSCI.0801-18.2018

Figure Lengend Snippet: Synaptic TrkB activation depends on ERα and β1 integrin, and is required for LTP in females. A, Deconvolved image shows punctate localization of pTrkB Y515 (red) and PSD-95 (green); yellow indicates double labeling (arrow). Scale bar, 2 μm. Line graph shows that TBS, compared with LFS, caused a greater rightward skew in the density frequency distribution for pTrkB-IR colocalized with PSD-95 (p < 0.0001, F(19,342) = 10.44), and MPP substantially reduced this effect (p < 0.0001, F(19,342) = 10.66; n = 10/group). Right, The percentage of double-labeled synapses with dense pTrkB-IR (≥90 units) was elevated after TBS in vehicle-treated, but not in MPP-treated, female slices (group means normalized to the LFS group mean; p = 0.0025, F(2,29) = 7.55; post hoc tests: LFS vs TBS, *p < 0.05; TBS vs TBS + MPP, ##p < 0.01). B, In slices from male rats, TBS increased both (left) the rightward skew in the synaptic pTrkB-IR density frequency distribution (vs LFS, p < 0.0001; F(19,342) = 6.794) that was not influenced by MPP (p = 0.939; F(19,342) = 0.549, n = 10/group) and (right) the percentage of PSD-95-IR synapses associated with dense pTrkB-IR (≥90 units), also not influenced by MPP (p = 0.009, F(2,29) = 5.68; Bonferroni's post-test: **p < 0.01 vs LFS). C, TrkB blocker ANA-12 (750 nm) disrupted the stabilization of CA1 LTP in female slices (p < 0.0001, t(10) = 8.36; n = 6/group). D, S-C TBS produced a marked increase in the percentage of PSDs associated with dense pTrkB-IR in wild-type mice but not in β1 integrin cKOs (p < 0.0001, F(3,36) = 25.55; post hoc tests: LFS vs TBS for wild types, ***p < 0.0001; LFS, n = 9; TBS, n = 8; LFS vs TBS for β1 cKOs, n.s.; n = 10/group). E, Image shows pCofilin-IR colocalized with PSD-95 in female CA1 SR. Right, In females, S-C TBS increased the rightward skew in the density frequency distribution for synaptic pCofilin (relative to LFS; p < 0.0001, F(19,399) = 7.69; LFS, n = 12; TBS, n = 11). F, The selective ROCK inhibitor H1152 (100 nm, 160 min) blocked S-C LTP in female slices (p = 0.0013, t(9) = 4.57; veh,, n = 6; H1152, n = 5).

Article Snippet: Primary antisera cocktails included rabbit antisera to pTrkB Y515 ( Wang et al., 2016a ; 1:500; catalog #NB100-92656, Novus Biologicals; RRID: AB_1218205 ), the activated conformation of β1 integrin ( Wang et al., 2016a ; 1:400; catalog #MAB2259Z, EMD Millipore; RRID: AB_94616 ), pFAK Y397 ( Bock and Herz, 2003 ; 1:500; catalog #44-624G, ThermoFisher Scientific; RRID: AB_2533701 ), pCofilin Ser3 ( Lauterborn et al., 2017 ; 1:500; catalog #ab12866, Abcam; RRID: AB_299488 ), pERK1/2 Thr202/Tyr204 ( Seese et al., 2014 ; 1:500; catalog #4370, Cell Signaling Technology; RRID: AB_2315112 ), phosphorylated (p) Src Tyr418 ( Chen et al., 2010 ; 1:500; catalog #44-660G, Thermo Fisher Scientific; RRID: AB_1500523 ), ERα (1:700; catalog #sc-542, Santa Cruz Biotechnology; RRID: AB_631470 ), or GPER1 (1:1000; catalog #ab39742, Abcam; RRID: AB_1141090 ) in combination with mouse anti-postsynaptic density-95 [PSD-95; 1:1000; catalog #MA1-045, Thermo Fisher Scientific (RRID: AB_325399 ); or catalog #ab12093, Abcam (RRID: AB_298846 )], or mouse anti-ERβ (1:700; catalog #sc-390243, Santa Cruz Biotechnology; RRID: AB_2728765 ) in combination with goat anti-PSD-95 (1:1000; catalog #ab12093, Abcam; RRID: AB_298846 ).

Techniques: Activation Assay, Labeling, Produced